【摘要】 目的:构建靶向肝脏的小凹蛋白-1 caveolin-1基因重组腺病毒并对该表达体系进行鉴定。方法:根据GeneBanks中caveolin-1的序列设计引物,以大鼠肝组织提取总RNA为模板,PCR扩增caveolin-1,并将caveolin-1序列克隆至pAdTrack-CMV质粒中,再与pAd-easy质粒进行同源重组构建重组腺病毒载体,利用脂质体法于293细胞中进行包装重组腺病毒,RT-PCR检测caveolin-1基因的转录,Westernblotting检测caveolin-1蛋白的表达情况。结果:PCR及Western blotting检测结果表明成功构建出caveolin-1基因重组腺病毒。结论:成功构建出caveolin-1基因重组腺病毒,为进一步研究caveolin-1基因在肝纤维化及肝硬化疾病中的作用机制打下实验基础。更多还原

【Abstract】 Objective:To construct and identify the target liver adenovirus expressing caveolin-1.Methods:According to Gene Banks Sequence, the primers of caveolin-1 sequence were designed, genomic RNA from ratliver tissue was extracted as a template, caveolin-1 was amplified by PCR, and the sequence of caveolin-1 wascloned into pAd Track-CMV plasmid, and homogonously recombined with pAd-easy plasmid to obtain therecombinant adenoviral vector. Recombination adenovirus was produced in 293 cells by lipofectamine method,transcription of caveolin-1 and expression of caveolin-1 were detected by RT-PCR and Western blotting,respectively.Results: Detection results of PCR and Western blotting showed that expressing caveolin-1recombination adenovirus were prosperity constructed.Conclusion: Successfully constructed caveolin-1recombination adenovirus helps further study of the effects of caveolin-1 in liver fibrosis and liver cirrhosis.更多还原